Cell and tissue samples are treated with lysates containing protease inhibitors and phosphatase inhibitors to prevent the enzymes in the sample from being released to hydrolyze or dephpsphprylate protein due to the disruption of the membrane system.
Sulfhydryl compounds react with 5,5' -dithiobis (2-nitrobenzoic acid) under neutral or alkaline conditions to produce a yellow product which have a maximum absorption peak at 412 nm. Measure the OD value and calculate the total mercapto content indirectly.
The superoxide anion free radical (O2-) can be produced by xanthine and xanthine oxidase reaction system, O2- oxidize hydroxylamine to form nitrite, it turn to purple under the reaction of developer. When the measured samples containing SOD, the SOD can specifically inhibit superoxide anion free radical (O2-). The inhibitory effect of SOD can reduce the formation of nitrite, the absorbance value of sample tube is lower than control tube. Calculate the SOD of sample according to the computational formula.
The activity of SOD was measured by WST-1 method in this kit and the principles of the WST-1 is as follows. Xanthine Oxidase (XO) can catalyze WST-1 react with O2.- to generate a water-soluble formazan dye. SOD can catalyze the disproportionation of superoxide anions, so the reaction can be inhibited by SOD, and the activity of SOD is negatively correlated with the amount of formazan dye. Therefore, the activity of SOD can be determined by the colorimetric analysis of WST-1 products.
Transglutaminases (TG) catalyze the transfer of acyl groups between glutamine residues and lysine residues or primary amines. TG-mediated protein crosslinking is essential for a variety of biological processes and diseases. TG is also widely used as a food processing aid in meat, fish, dairy and grain products. The detection principle of this kit is: TG catalyzes the acyl transfer between glutamine residues and primary amines, the product forms a chromogenic substance with the chromogenic agent, and determines the activity of TG in the sample by measuring the absorbance change at 525 nm.
Trehalose, a non-reducing disaccharide composed of two glucose molecules, is widely found in lower plants, invertebrates and microorganisms. Trehalose is the main energy storage substance for many insects and fungi. In addition, trehalose has a non-specific protective effect on organisms and biological macromolecules. This special property makes it widely used in many fields such as cosmetics, food industry, medicine and agriculture. This kit specifically hydrolyzes trehalose into two molecules of glucose by trehalose enzyme, and then detects the glucose content by the GOD-POD method. Control well is set up to eliminate the influence of the glucose background value, thereby determining the trehalose content in the sample. It has the characteristics of strong specificity and high sensitivity.
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